I have used Qiagen kit for RNA extraction from 100 mg normal brain tissue ( human). I am getting 1600 (approx) ng/ul. It was eluted in 25ul nuclease free water. Tissue was taken from a distant site ( cortex) from a low grade astrocytoma patient. Histopathological examination of part of this tissue showed normal morphology. I still have a doubt if this is normal owing to high RNA yield. Checked the quality on gel as well. It was good.
Based on your experience could you kindly suggest if I can use this sample as a control?
Hi Ira,
Good question. I assume you are using fresh tissues right?. Your RNA yield is very good and its pretty high. Do you need to convert them to cDNA as well. Make sure you use >1000ng/ul for cDNA conversions. We use 5 to 30 mg tissues for RNA extractions and we get ~ 100ng/ul. So your concentrations are in right place. Good luck with your future research.
I use exactly the similar kit here in my lab. The RNA concentration you received is an ideal amount for 100mg tissue. I use 5-20 mg tissue and my concentration is approximately 100-200ng/micro liter. You can probably use less amount of starting material as the filters may not be capable to bind with these many RNA molecules. Please make sure you use an appropriate cDNA conversion kit too.
Best Wishes
I usually get high RNA yield in tumor tissues ( GBM and astrocytoma). Hence when I got this yield in supposedly normal tissue I got little wary. As I am getting > 1000ng/ul -3000ng/ul concentration in tumor (brain) tissue. In one reference from miltenyi biotec average RNA yield from 30mg normal brain tissue was mentioned as 1.7ug. So in my case it should be about 5-6 ug in total. But I am getting a total yield of >40ug. If this is not normal then I may not get any different profile from rest of the tumors in my study.
Are you using postmortem brain tissue as control or commercially available RNA?
My experience is based on Trizol extraction from human developing brain (Medulla oblongata).
From a tissue of 95mg I yield 1060ng/ul in a volume of 50ul.
Hi guys,
I am going to probably isolate RNA from human brain tissues. I would like to sue Qiagen kit since I can also isolate protein and gDNA at the same time.
Have any of you published any of your results? Can you share your publication?
Thank you!
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